Zīst/shināsī-i Giyāhī-i Īrān (Mar 2014)
Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
Abstract
Transient gene expression is fast, easy and not influenced by positional effects that potentially affect on gene expression levels in stable gene transformation. Transient expression can be applicable using agroinfilteration, biolistic and viral vectors. Agrobacterium mediated transient expression have been shown as an efficient and versatile method for analyzing transgene expression, gene scilencing, host-pathogen interactions, protein-protein interaction, and cis-element/transfactor interaction. A control vector consists of an interon containing reporter gene under control of a common promoter is often required for Cis-acting elemen analysis to standardize the variation. This study was carried out to construct a control vector based on two parental binary vectors, pGPTV and pCAMBIA3301. The constructed vector, pGCGi had an interon containing GUS reporter gene under control of the full sequence of CaMV 35S promoter. The GUS staining results revealed that the GUS intron containg gene cannot produce an active B- glucronidase enzyme in agrobacterial cells. The functional gene expression analysis of the new vector was done using agroinjection and prticle delivery system in tobacco and onion eoidermal cells respectively. The histochemical GUS staining results showed pGCGi could express the reporter gene in plant cells and culd be used as control vector in transient expression experiments.
